Reagents for flow cytometry

  1. PBS 10 X (Stock)

  2. PBSA 1 X (Working)

  3. Lysing solution 10 X & 1 X

  4. PBS-PFA

1 liter stock of 10x PBS can be prepared by dissolving

  • 7.2 g NaCl,

  • 1.32 g Na2HPO4 · 2H2O

  • 7.52 g KH2PO4

  • Make upto 1 L by distilled water.

  • pH 7.2 ± 0.2, Store 2-80C in tightly closed bottle

  • Dil stock 1 in 10 to a total vol that you would consume in a weak for 1x PBS working

1 liter stock of 1x PBSA can be prepared by dissolving

  • 10g BSA

  • 2.5g Sodium Azide

  • Make upto 1 L by 1x PBS.

  • pH 7.2 ± 0.2, Store 2-80C in tightly closed bottle

1 litre of Lysing solution stock (10X)

  • 89.9g NH4Cl

  • 10.0g KHCO3

  • 3.7g EDTA Na4

  • DW to 1 lit.

  • pH 7.2 ± 0.2, Store 2-80C in tightly closed bottle

100ml Lysing solution working (1X)

  • 90 ml DW + 10 ml 10X stock

  • Mix well. Store at RT. Discard after 3 days

PBS – PFA (0.5%)

  • PFA 0.5g in 100 ml of 1 X PBS (with azide)

  • Dissolve by heating to 65-700C (hot plate with magnetic stirrer is best). Must keep flask/beaker covered tightly with AL foil while heating.

  • Filter solution though a 0.2 μ suction filter before use esp for minimal residual disease work.

  • Store at 40C for upto 2 weaks

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